Cloning of genes which regulate mortality

克隆调节死亡率的基因

• 批准号: 04671353
• 负责人: IDE Toshinori
• 金额: $ 1.34万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671353/
• 关键词: Fibroblasts; Senescence; CDNA library; Cytokines; Interferon; Growth repression; サイトカン; ヒト線維芽細胞; T抗原; 分裂寿命; 培養細胞

human normal fibroblasts have a finit proliferative lispan (normal). Cancer cells are generally infinit lifespan (immortal). The purpose of the present project is to search for genes which participate in growth repression at the end of the proliferative lifespan of normal cells. Subtraction cDNA libraies were prepared which is expected to enrich cDNAs that highly expressed in senescent cells. Clones were selected by serial differential plaque hybridization screening from the library. Expression of these genes was confirmed by northern blot. Nine clones were finally selected. By sequencing and homology search of these clones, 4 clones were found to be unreported sequences, 4 clones to be highly homologous to known genes, one of which was interferon (IFN)-inducible gene, and 1 clone to be a partial sequence of known gene, IFN-inducible gene 6-16. Considering that it was unexpectedly high incidence of IFN-inducible gene or its homologue and that IFN has growth repressive effect, IFN can be a candidate of genes which participate on growth supression of senescent cells. IFN-beta, not IFN-alpha and IFN-gamma, was produced at the end of the lifespan where high level induction occurred in three other IFN-inducible genes which was reduced by anti-IFN-beta antibody treatment.

人正常成纤维细胞有一个有限的增殖性口吃(正常)。癌细胞通常是无限寿命(永生的)。本项目的目的是寻找在正常细胞增殖寿命结束时参与生长抑制的基因。消减文库的制备有望丰富衰老细胞中高表达的cDNA。用连续差示斑块杂交方法从文库中筛选克隆。Northern印迹分析证实了这些基因的表达。最终选出了9个无性系。通过测序和同源性搜索，发现4个克隆为未报道序列，4个克隆与已知基因高度同源，其中1个为干扰素诱导基因，1个克隆为已知基因的部分序列，即干扰素诱导基因6-16。考虑到干扰素诱导基因或其同源物的高发生率，以及干扰素具有生长抑制作用，干扰素可能是参与衰老细胞生长抑制的候选基因。干扰素-β是在生命结束时产生的，而不是干扰素-α和干扰素-γ，在生命结束时，另外三个干扰素诱导基因发生了高水平的诱导，而这些基因被抗干扰素-β抗体处理而减少。

项目成果

井出利憲: "老化のメカニズムと制御" アイピーシー社, 465 (1993)

Toshinori Ide：“衰老的机制和控制”IPC Publishing，465（1993）

Eiji Hara, Tomoko Ymaguchi, Susumu Nakata, Toshinori Ide, Kinichiro Oda.: "Senescence and immortalization of human fibroblasts." Soshikibaiyo Kenkyuu. 11. 43-48 (1992)

Eiji Hara、Tomoko Ymaguchi、Susumu Nakata、Toshinori Ide、Kinichiro Oda.：“人类成纤维细胞的衰老和永生化。”

Eiji Hara, Tomoko Ymaguchi, Hidetoshi Tahara, Naohiro, Tsuyama, Hiromichi Tsurui, Toshinori Ide, and Kinichiro Oda.: "DNA-DNA subtractive cDNA cloning using oligo(dT)(30)-Latex and PCR-identification of cellular genes which are overexpressed in senescent

Eiji Hara、Tomoko Ymaguchi、Hidetoshi Tahara、Naohiro、Tsuyama、Hiromichi Tsurui、Toshinori Ide 和 Kinichiro Oda.：“使用oligo(dT)(30)-Latex 进行 DNA-DNA 消减 cDNA 克隆和 PCR 鉴定细胞基因，这些细胞基因是

原英二: "ヒト線維芽細胞の老化(不死化)関連遺伝子" 組織培養研究. 11. 43-48 (1992)

Eiji Hara：“人类成纤维细胞中的衰老（永生）相关基因”组织培养研究 11. 43-48 (1992)。

田原栄俊: "Preparation of subtractive cDNA library enriched in cDNAs which expressed at high level" Biochem.Biophys.Res.Commun.(印刷中). (1994)

Hidetoshi Tahara：“富含高水平表达的 cDNA 的消减 cDNA 文库的制备”Biochem.Biophys.Res.Commun.（出版中）。